EP1414937B1 - Extraits de houblon, procede destine a leur preparation et leur utilisation - Google Patents

Extraits de houblon, procede destine a leur preparation et leur utilisation Download PDF

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Publication number
EP1414937B1
EP1414937B1 EP02758454A EP02758454A EP1414937B1 EP 1414937 B1 EP1414937 B1 EP 1414937B1 EP 02758454 A EP02758454 A EP 02758454A EP 02758454 A EP02758454 A EP 02758454A EP 1414937 B1 EP1414937 B1 EP 1414937B1
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Prior art keywords
extract
hop
prenylnaringenin
solvent
group
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EP02758454A
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German (de)
English (en)
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EP1414937A1 (fr
Inventor
Clemens Erdelmeier
Egon Koch
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Dr Willmar Schwabe GmbH and Co KG
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Dr Willmar Schwabe GmbH and Co KG
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    • C—CHEMISTRY; METALLURGY
    • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12C—BEER; PREPARATION OF BEER BY FERMENTATION; PREPARATION OF MALT FOR MAKING BEER; PREPARATION OF HOPS FOR MAKING BEER
    • C12C9/00—Methods specially adapted for the making of beerwort
    • C12C9/02—Beerwort treatment; Boiling with hops; Hop extraction
    • C12C9/025—Preparation of hop extracts ; Isomerisation of these extracts; Treatment of beerwort with these extracts; Surrogates of the hop
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K36/00—Medicinal preparations of undetermined constitution containing material from algae, lichens, fungi or plants, or derivatives thereof, e.g. traditional herbal medicines
    • A61K36/18—Magnoliophyta (angiosperms)
    • A61K36/185—Magnoliopsida (dicotyledons)
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P13/00—Drugs for disorders of the urinary system
    • A61P13/08—Drugs for disorders of the urinary system of the prostate
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P15/00—Drugs for genital or sexual disorders; Contraceptives
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P19/00—Drugs for skeletal disorders
    • A61P19/08—Drugs for skeletal disorders for bone diseases, e.g. rachitism, Paget's disease
    • A61P19/10—Drugs for skeletal disorders for bone diseases, e.g. rachitism, Paget's disease for osteoporosis
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P25/00—Drugs for disorders of the nervous system
    • A61P25/28—Drugs for disorders of the nervous system for treating neurodegenerative disorders of the central nervous system, e.g. nootropic agents, cognition enhancers, drugs for treating Alzheimer's disease or other forms of dementia
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P5/00—Drugs for disorders of the endocrine system
    • A61P5/24—Drugs for disorders of the endocrine system of the sex hormones
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P5/00—Drugs for disorders of the endocrine system
    • A61P5/24—Drugs for disorders of the endocrine system of the sex hormones
    • A61P5/30—Oestrogens
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P9/00—Drugs for disorders of the cardiovascular system
    • C—CHEMISTRY; METALLURGY
    • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12C—BEER; PREPARATION OF BEER BY FERMENTATION; PREPARATION OF MALT FOR MAKING BEER; PREPARATION OF HOPS FOR MAKING BEER
    • C12C3/00—Treatment of hops
    • C12C3/04—Conserving; Storing; Packing
    • C12C3/08—Solvent extracts from hops
    • C12C3/10—Solvent extracts from hops using carbon dioxide

Definitions

  • the present invention relates to hop extracts, methods for their production and the use of hop extracts for the prophylaxis and therapy of disease states that by a lack of estrogens or by dysregulations the sex hormone metabolism, in particular the Estrogen metabolism.
  • Hops have had a medicinal substance for a long time Meaning as a mild sedative in folk medicine. Responsible for this effect are probably the oxidation-sensitive ⁇ - and ⁇ -bitter-acids.
  • Ingredients have been more recently Radical scavenging properties as well as lipid peroxidation inhibitory properties (M. Tagashira et al., Biosci. Biotech. Biochem. 59, 740-742 (1995)).
  • EP 0 677 289 A2 are also pharmaceutical Compositions for the treatment of osteoporosis described the compounds from the group of ⁇ -bitter-acids and the ⁇ -iso-bitter acids.
  • xanthohumol 1 inhibits bone loss. Its use as a therapeutic for osteoporosis is described in EP 0 679 393 B1. Although the inventors postulate estrogenic properties of xanthohumol, they are not demonstrated. On the contrary, SR Milligan et al. (Pharm. Pharmacol. Lett., 7: 83-86 (1997)) clearly indicates that the osteoporosis-inhibiting activity of xanthohumol is due to an estrogenic effect, since it does not exert any similar activity on the human endometrial carcinoma cell line Ishikawa or in a yeast.
  • xanthohumol 1 and isoxanthohumol 4 bind with comparable activity to the estrogen receptors alpha and beta.
  • WO 83/00701 A1 discloses a method for obtaining estrogen-active substances from hops claimed thereby characterized in that first a carbon dioxide extract Hops prepared with the addition of water as an entraining agent and then from it the estrogen-active substances by means of ether extraction or chromatographic methods to be obtained. Furthermore, the use of these substances as Additive to animal feed, for cosmetic products or as Bath additive claimed. About the nature of this estrogen-active substances are given no information.
  • the present invention is based on the object To provide plant extracts useful for the manufacture of Medicines for the prophylaxis and therapy of Disease conditions that are due to a lack of Estrogens or by dysregulations of sex hormone metabolism, especially the estrogen metabolism, caused.
  • Another object of the invention is the provision a process for producing such extracts and these comprehensive pharmaceutical preparations for treatment the above disease states are suitable.
  • hops extract according to the invention according to claim 1 and 2 the method according to the Claims 3-12, the pharmaceutical preparation according to Claim 13 and the use of the extracts or the Pharmaceutical preparation according to claims 14-16 solved.
  • the present invention is based inter alia on the surprising finding that from hop drug after removal of lipophilic and hydrophilic fiber extracts which are the phloroglucinolartigen Hop bitter acids still included, at the same time free and / or bound chalcones and flavones such as xanthohumol, Isoxanthohumol and 6- and 8-prenylnaringenin in contrast enriched form included.
  • FIG. 1 illustrates the dependence of the concentration the analyzed ingredients of the temperature of the Wasservorextralement.
  • Such an extract can be selected from one to several times extraction with a C 5 -C 7 alkane or with supercritical CO 2 (degreasing stage), subsequent extraction of the remaining drug residue with water and subsequent extraction of the remaining drug residue with a medium polar solvent selected from Group consisting of alcohols, aqueous alcohols, ketones, aqueous ketones, esters and optionally subsequent liquid-liquid distribution can be obtained.
  • a medium polar solvent selected from Group consisting of alcohols, aqueous alcohols, ketones, aqueous ketones, esters and optionally subsequent liquid-liquid distribution
  • the hop bitters are not completely, but only partially in the lipophilic extract, while on the other hand, the chalcone and flavones remain almost completely in the drug residue in the water extraction.
  • this makes it possible to obtain a hop extract containing all pharmacologically relevant ingredients (bitter acids, chalcones, flavones) in a balanced ratio.
  • this extract is ideally useful in disease states caused by a lack of estrogens or other hormonal dysregulations.
  • the hops extracts according to the invention are for prophylaxis and Treatment with menopause or postmenopause symptoms associated with women, with the Symptoms including hot flushes, depression, Anxiety, mental confusion, insomnia as well serious post-menopausal related Health problems such as osteoporosis, cardiovascular disease, Stroke, dementia and tumors include.
  • Other disorders that indicate dysregulation of sex hormone metabolism and with the Extract according to the invention can be treated, e.g. Amenorrhoea, anovulatory cycles, menorrhagia, premenstrual symptoms and postpartum depression.
  • these extracts can be used for treatment Sex hormone-dependent diseases used in men be such. benign prostatic hyperplasia or Prostate cancer.
  • Figure 2 shows the activity of a comparative extract and of two hop extracts according to the invention in a yeast reporter gene assay.
  • a hop extract is compared with one conventional, especially aqueous-alcoholic extracts increased content of free and / or bound chalcones and Flavones, especially 6- and 8-prenylnaringenin, xanthohumol and Isoxanthohumol provided simultaneously ⁇ and possibly ⁇ -bitter-acids (Humulon or Lupulon and its Derivatives).
  • the ratio of drug to solvent is at each Extraction step in the range of about 1: 7 to about 1:12.
  • the extraction with a C 5 -C 7 alkane or supercritical CO 2 in step (a) is preferably carried out once, twice or three times, in particular three times.
  • the extraction with supercritical CO 2 is particularly preferred.
  • the C 5 -C 7 alkane in step (a) is preferably a C 5 -C 7 n-alkane selected from the group consisting of n-pentane, n-hexane and n-heptane, with n-heptane being most preferred ,
  • the extraction in step (b) is preferably at 90 ° C, carried out, wherein the extraction time one or more Hours can be.
  • the solvent in step (c) is preferably selected consisting of the group 74 to 99% (g / g) Methanol or 60 to 99% (g / g) acetone and 92% (g / g) ethanol.
  • the hops (dry) extract according to the invention is characterized by a content of ⁇ -bitter-acids of at least 0.5%, preferably at least 0.8% and in particular at least 1%, of xanthohumol of at least 2%, preferably at least 3% and in particular at least 4% and at prenylated flavones of at least 0.5%, preferably at least 0.7%.
  • the prenylated flavones include preferably 6-prenylnaringenin, 8-prenylnaringenin and Isoxanthohumol.
  • Xanthohumol is within the meaning of the present The invention does not belong to the prenylated flavones.
  • the Percentages refer to the weight of the Hops dry extract.
  • the extracts obtained can be used together with usual pharmaceutically acceptable excipients to pharmaceutical Preparations such as capsules, film-coated tablets and dragees are processed.
  • pharmaceutical excipients conventional fillers, binders, blasting agents, lubricants and coating agents for film-coated tablets and dragees as well as oils and fats as Fillers used for soft gelatin capsules.
  • the extracts according to the invention can be used for the prophylaxis and Therapy of disease states used by a lack of estrogen or other hormonal Dysregulations are caused, in particular climacteric complaints, sex hormone dependent Cancers, benign prostatic hyperplasia, osteoporosis, Alzheimer's disease and cardiovascular disease.
  • the extracts according to the invention may be used in particular for Prophylaxis and therapy of breast cancer, uterine cancer and Prostate cancer can be used.
  • the dosage of the extracts according to the invention is in the range from 0.005 g to 2 g of extract 1 to 4 times a day, preferably in the range of 0.02 g to 1 g 1 to 2 times a day.
  • the Dosage in individual cases depends on the clinical picture and the individual circumstances of the patient and can of the attending specialist according to the respective Be adapted to needs.
  • the hop drug (variety "Hallertauer Magnum") were mixed with 500 g of 96% (g / g) of ethanol and comminuted with the Ultraturrax. It was extracted at 60 ° C for 1 h. It was then filtered through a Seitz 1500 filter. The drug was extracted 2 more times the same way.
  • the combined extract solutions were freed from ethanol on a rotary evaporator and dried overnight in a vacuum oven at 50 ° C. From the dry matter, the content of characteristic ingredients is determined by the following HPLC method. This HPLC method is used to determine the ingredients in the other examples. pillar LiChrospher 100 5 ⁇ m.
  • Example 1a Production of a Hop Extract (Extraction with CO 2 and Water Pre-extraction at 90 ° C.)
  • the water extract was filtered off through a Seitz Supra Filter 1500.
  • the still slightly damp drug residue was then extracted with 2 times 800 g of 92% (g / g) of ethanol in each case 1 hour at 60 ° C. It was then filtered off on Seitz Supra 1500 and the extract solution on a rotary evaporator at a water bath temperature of 55-65 ° C freed of ethanol and dried in an oven at 60 ° C.
  • Example 1b Preparation of a Hop Extract (Extraction with CO 2 and Water Pre-extraction at 90 ° C.)
  • a hop drug (variety "Hallertauer Magnum"), which had been previously pre-extracted with supercritical CO 2 (conditions: grinding to 10 mm grain size, extraction with CO 2 at 250 bar / 50 ° C, separation of the extract in a yield of 30%) were extracted with 6 kg of water first for 5 min. on Ultra-Turrax, then with stirring for 1 hour at 90 ° C. Subsequently, the water extract was filtered off through a Seitz Supra Filter 1500. The still slightly damp drug residue was then extracted with 2 times 5 kg of 92% (g / g) of ethanol each at 60 ° C for 1 hour.
  • a hop drug variety "Hallertauer Magnum”
  • a hop drug (variety "Hallertauer Magnum"), which had been previously pre-extracted with supercritical CO 2 (conditions: grinding to 10 mm grain size, extraction with CO 2 at 250 bar / 50 ° C, separation of the extract with a yield of 30%) were extracted with 964 g of water first for 5 min. on Ultra-Turrax, then with stirring for 1 hour at 60 ° C. Subsequently, the water extract was filtered off through a Seitz Supra Filter 1500. The still slightly damp drug residue was then extracted with 2 times 800 g of 92% (g / g) of ethanol for 5 min. First on Ultra-Turrax, then with stirring for 1 hour at 60 ° C extracted.
  • a hop drug variety "Hallertauer Magnum”
  • hop drug variety "Hallertauer Magnum”
  • hop drug variety "Hallertauer Magnum”
  • n-heptane After filtering off the heptane extract solution on Seitz Supra 1500 was extracted a second time in the same manner. Thereafter, the remaining drug residue in the vacuum oven was freed from heptane.
  • the dry drug residue (205 g) was then added with 12 times the weight of water and kept at 90 ° C for 1 hour. It was then filtered off again and the still slightly damp drug residue with 10 times the amount by weight 92% (g / g) of ethanol with stirring twice at 60 ° C extracted.
  • results shown graphically in FIG. 1 show a clear dependence of the concentration of analyzed prenylated ingredients from temperature the water pre-extraction.
  • a comparison extract and an extract according to the invention for interactions with the human estrogen receptor alpha (ER- ⁇ ) or beta (ER- ⁇ ) was carried out.
  • Radiolabeled oestradiol is first bound to the human estrogen receptor and then treated with the test substance to be investigated. A proportion of labeled estradiol corresponding to the estrogenic potency of the sample is thereby displaced. Excess estradiol is washed out after binding the complex to hydroxyapatite.
  • the estrogen receptors ER- ⁇ and ER- ⁇ were purchased commercially as recombinant human receptors.
  • test mixtures each consisted of 1000 ⁇ l of TEDG buffer (10 mM Tris, 1.5 mM EDTA, 10% glycerol, pH 7.5), 5 ⁇ l receptor (200 nM), 10 ⁇ l 3H-estradiol and 10 ⁇ l ethanol (control value), 10 ⁇ l of diethylstilestrol (100 mM, positive control) or 10 ⁇ l of extract or extract.
  • TEDG buffer 10 mM Tris, 1.5 mM EDTA, 10% glycerol, pH 7.5
  • 5 ⁇ l receptor 200 nM
  • 10 ⁇ l 3H-estradiol and 10 ⁇ l ethanol control value
  • 10 ⁇ l of diethylstilestrol 100 mM, positive control
  • extract or extract 10 ⁇ l of extract or extract.
  • the batches are gently mixed and incubated for about 16 hours at room temperature in the dark. After incubation, 250 ⁇ l of hydroxyapatite (HAP) is added to adsorb
  • the precipitate is spun down sharply at 10,000 rpm for a few seconds and the supernatant is pipetted off.
  • the pellet is washed three times with 1000 .mu.l of TEDG buffer and mixed with 1000 .mu.l of ethanol for measurement, slurried and transferred to a scintillation vial. After adding 9 ml of scintillator fluid (Ready Safe, Beckmann), a measurement is made over the entire 3 H window in a Beckmann Beta Counter.
  • the binding capacities of the test substances are characterized by determining the ED 50 values from the dose-response curves of oestradiol displacement.
  • the results are summarized in Tab. 1 and demonstrate potent interactions with both estrogen receptors for all ingredients tested.
  • the extract of the invention proved to be much more effective than would be expected from the activities of the individual ingredients.
  • the reference extract showed an activity at both receptors that was at least 10-fold lower than that of the extract according to the invention.
  • the testing of extracts for estrogenic properties was also carried out with a reporter gene assay using yeast cells (Saccharomyces).
  • yeast cells Sacharomyces
  • the cells are stably transfected with the human ⁇ -estrogen receptor and an expression plasmid containing an estrogen response element and the gene for the enzyme ⁇ -galactosidase. All samples were dissolved in DMSO at a concentration of 20 mg / ml and, after dilution with DMSO in the ratio 1/10, 1/100 or 1/1000 in a volume of 1 .mu.l to 100 .mu.l culture medium in 96-well flat-bottomed Given microtiter plates.

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Claims (15)

  1. Procédé d'obtention d'un extrait de houblon, comprenant les étapes consistant à :
    (a) extraire une ou plusieurs fois une drogue de houblon avec un alcane en C5-C7 ou du CO2 supercritique et séparer le résidu de drogue de la solution d'extraction ;
    (b) extraire une ou plusieurs fois le résidu de drogue de l'étape (a) avec de l'eau à une température située dans la plage de 60 à 95°C et séparer le résidu de drogue ;
    (c) extraire une ou plusieurs fois le résidu de drogue de l'étape (b) avec un solvant choisi dans le groupe constitué de 80 à 96 % (g/g) d'éthanol, de méthanol, de méthanol aqueux, d'acétone, d'acétone aqueuse et d'acétate d'éthyle, ainsi que filtrer la solution d'extraction obtenue ; et
    (d) éliminer le solvant des solutions d'extrait réunies, obtenues à l'étape (c), et sécher le résidu obtenu.
  2. Procédé selon la revendication 1, sachant que, à l'étape (a), on extrait une fois, deux fois ou trois fois.
  3. Procédé selon l'une des revendications 1 ou 2, sachant que le solvant à l'étape (a) est choisi dans le groupe constitué du n-pentane, n-hexane et n-heptane.
  4. Procédé selon la revendication 3, sachant que le solvant à l'étape (a) est le n-heptane.
  5. Procédé selon l'une des revendications 1 à 4, sachant que l'extraction à l'étape (b) se fait à environ 90°C.
  6. Procédé selon l'une des revendications 1 à 5, sachant que le solvant à l'étape (c) est choisi dans le groupe constitué de 92 % (g/g) d'éthanol, 74 à 99 % (g/g) de méthanol et 60 à 99 % (g/g) d'acétone.
  7. Procédé selon la revendication 1, sachant que le solvant à l'étape (a) est le n-heptane et le solvant à l'étape (c) est 92 % (g/g) d'éthanol.
  8. Procédé selon la revendication 1, sachant que le solvant à l'étape (a) est du CO2 supercritique et le solvant à l'étape (c) est 92 % (g/g) d'éthanol.
  9. Extrait de houblon pouvant être obtenu d'après le procédé selon l'une des revendications 1 à 8, caractérisé par une teneur en acides amers alpha d'au moins 0,5 %, en xanthohumol d'au moins 2 % et en flavones prénylées choisies dans le groupe composé de la 6-prénylnaringénine, 8-prénylnaringénine et de l'isoxanthohumol d'au moins 0,5 %.
  10. Extrait de houblon selon la revendication 9, caractérisé par une teneur en acides amers alpha d'au moins 0,8 %, en xanthohumol d'au moins 3 % et en flavones prénylées choisies dans le groupe composé de la 6-prénylnaringénine, 8-prénylnaringénine et de l'isoxanthohumol d'au moins 0,7 %.
  11. Composition pharmaceutique, comprenant un extrait de houblon, caractérisé par une teneur en acides amers alpha d'au moins 0,5 %, en xanthohumol d'au moins 2 % et en flavones prénylées choisies dans le groupe composé de la 6-prénylnaringénine, 8-prénylnaringénine et de l'isoxanthohumol d'au moins 0,5 %, et d'autres excipients pharmaceutiquement acceptables.
  12. Composition pharmaceutique selon la revendication 11, comprenant un extrait de houblon, caractérisé par une teneur en acides amers alpha d'au moins 0,8 %, en xanthohumol d'au moins 3 % et en flavones prénylées choisies dans le groupe composé de la 6-prénylnaringénine, 8-prénylnaringénine et de l'isoxanthohumol d'au moins 0,7 %.
  13. Utilisation d'un extrait de houblon, tel qu'il est défini dans la revendication 9 ou 10 ou d'une composition pharmaceutique d'après la revendication 11 ou 12 pour la préparation d'un médicament destiné à la prophylaxie et la thérapie des états pathologiques, provoqués par une déficience en oestrogènes ou par une dérégulation du métabolisme des hormones sexuelles, en particulier du métabolisme des oestrogènes, choisis dans le groupe constitué des troubles climatériques, des hyperplasies de la prostate bénignes, des ostéoporoses, de la maladie d'Alzheimer et des affections cardio-vasculaires.
  14. Utilisation d'un extrait de houblon tel que défini dans la revendication 9 ou 10 ou d'une composition pharmaceutique d'après la revendication 11 ou 12 pour la préparation d'un médicament destiné à la prophylaxie des maladies cancéreuses dépendantes des hormones sexuelles.
  15. Utilisation selon la revendication 14, sachant que les maladies cancéreuses dépendantes des hormones sexuelles sont choisies dans le groupe constitué du cancer du sein, cancer de la prostate et cancer de l'utérus.
EP02758454A 2001-08-10 2002-08-09 Extraits de houblon, procede destine a leur preparation et leur utilisation Expired - Lifetime EP1414937B1 (fr)

Applications Claiming Priority (3)

Application Number Priority Date Filing Date Title
DE10139479 2001-08-10
DE10139479A DE10139479A1 (de) 2001-08-10 2001-08-10 Hopfenextrakte, Verfahren zu ihrer Herstellung und Verwendung
PCT/EP2002/008943 WO2003014287A1 (fr) 2001-08-10 2002-08-09 Extraits de houblon, procede destine a leur preparation et leur utilisation

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EP1414937A1 EP1414937A1 (fr) 2004-05-06
EP1414937B1 true EP1414937B1 (fr) 2005-06-08

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US (1) US20050042318A1 (fr)
EP (1) EP1414937B1 (fr)
JP (1) JP2004537604A (fr)
CN (1) CN1269942C (fr)
AT (1) ATE297460T1 (fr)
DE (2) DE10139479A1 (fr)
ES (1) ES2240787T3 (fr)
MX (1) MXPA04001226A (fr)
WO (1) WO2003014287A1 (fr)

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US7413758B2 (en) 2002-03-26 2008-08-19 Millercoors Llc Preparation of light stable hop products
US7258887B2 (en) 2002-03-26 2007-08-21 Miller Brewing Company Preparation of light stable hops
EP1360959A1 (fr) * 2002-05-10 2003-11-12 Schering Aktiengesellschaft Utilisation des 8-prénylflavanones pour la thérapie anti-angiogénèse et la thérapie fibrinolitique
DE10240065B4 (de) * 2002-08-30 2004-10-28 Hallertauer Hopfenveredlungsgesellschaft M.B.H. Verfahren zur Gewinnung von in Hopfen enthaltenem Xanthohumol und damit erhältlicher Xanthohumol-reicher Hopfenextrakt
DE10256031A1 (de) * 2002-11-30 2004-06-09 Nateco 2 Gmbh & Co. Kg Verfahren zur Herstellung eines Xanthohumol-angereicherten Hopfenextraktes und dessen Verwendung
US20070059393A1 (en) * 2003-04-08 2007-03-15 Kirin Beer Kabushiki Kaisha Composition for inhibition or prevention of bone density
EP1524269A1 (fr) * 2003-10-07 2005-04-20 Schering Aktiengesellschaft Utilisation de 8-prenylnaringening pour hormonotherapie substitutive
EP1543834A1 (fr) * 2003-12-16 2005-06-22 Biodynamics Production de d'extraits de houblon ayant une bioactivité oestrogénique et anti-proliférative
DE102004012830A1 (de) * 2004-03-16 2005-10-06 Dr. Willmar Schwabe Gmbh & Co. Kg Hopfenextrakte, Herstellung und Verwendung
US20070218155A1 (en) * 2004-08-20 2007-09-20 Kuhrts Eric H Methods and compositions for treating dyslipidaemia
ITMI20060122A1 (it) * 2006-01-25 2007-07-26 Polichem Sa Composizioni per uso vaginale
DE102006018988B3 (de) * 2006-04-25 2007-08-09 Nateco2 Gmbh & Co. Kg Verfahren zur Herstellung eines Xanthohumol-haltigen Pulvers hoher Reinheit und dessen Verwendung
EP2023747A2 (fr) * 2006-05-24 2009-02-18 DSMIP Assets B.V. 3,5-dihydroxy-2,4-cyclohexadiénones et leurs dérivés, compositions diététiques et pharmaceutiques les comprenant, et leurs utilistions
CN101161807B (zh) * 2006-10-11 2011-09-28 韩延欣 从啤酒花中萃取分离啤酒花浸膏和啤酒花精油的方法
JP2008214261A (ja) * 2007-03-02 2008-09-18 Sapporo Breweries Ltd 芳香性化合物、芳香性組成物、芳香性ホップ抽出物
US20080233221A1 (en) * 2007-03-23 2008-09-25 State Of Oregon Prostate cancer and benign prostatic hyperplasia treatments
CA2696311C (fr) * 2007-08-15 2013-07-16 Flaxan Gmbh & Co. Kg Extrait de houblon enrichi en xanthohumol
US8142821B2 (en) * 2007-08-15 2012-03-27 Flazan GmbH & Co. KG Xanthohumol-enriched hop extract
CN101574398B (zh) * 2008-05-09 2011-04-27 上海信谊百路达药业有限公司 啤酒花提取物及其制备方法与应用
FR2940089B1 (fr) * 2008-12-24 2011-03-18 Greenpharma Sas Procede de preparation d'un produit enrichi en flavonoides a partir d'un materiel vegetal
EP2392325A1 (fr) 2010-06-04 2011-12-07 Universitätsklinikum Münster Nouveaux composés pour la prévention et/ou le traitement de l'arthrose
ES2394350B1 (es) * 2011-06-16 2013-12-02 Universidad Autónoma de Madrid Procedimiento para la obtención de extracto de lúpulo y extracto obtenido
CN102911033A (zh) * 2011-08-02 2013-02-06 苏州宝泽堂医药科技有限公司 一种从啤酒花中制备黄腐酚的方法
RU2014108490A (ru) 2011-08-08 2015-09-20 Университет Медычны В Лодзи Полифенольная композиция, ее получение и применение
CN103371989A (zh) * 2012-04-24 2013-10-30 华东师范大学 黄腐酚在制备抑制破骨细胞分化和蚀骨功能的预防和/或治疗药物中的应用
WO2014025989A1 (fr) * 2012-08-10 2014-02-13 Eric Hauser Kuhrts Compositions et procédés de traitement de troubles de l'humeur ou d'une maladie ou lésion de la peau
CN103285087A (zh) * 2013-01-23 2013-09-11 中日友好医院 啤酒花有效部位用于制备预防及改善焦虑症及其临床症状的药物的应用
CN103156923A (zh) * 2013-01-23 2013-06-19 北京中医药大学 啤酒花有效部位用于制备预防和改善抑郁症及其临床症状药物的应用
EP2759295A1 (fr) * 2013-01-29 2014-07-30 Max-Planck-Gesellschaft zur Förderung der Wissenschaften e.V. Cellule de mousse spécifique de l'agoniste alpha du récepteur des oxystérols (LXR), inhibiteurs de SIRT1 ainsi qu'inhibiteurs de protéines p300 comme agents pharmaceutiquement actifs
CN104116022B (zh) * 2013-04-26 2016-08-10 中国科学院理化技术研究所 用于改善妇女更年期综合症的复合保健食品及其制备方法
PL223580B1 (pl) 2013-10-04 2016-10-31 Bioactive Tech Spółka Z Ograniczoną Odpowiedzialnością Sposób otrzymywania ksantohumolu o wysokiej czystości
CN104189086B (zh) * 2014-09-12 2017-02-08 吉林省中医药科学院 啤酒花天然药物组合物的制备方法及用途
AU2015379412A1 (en) * 2015-01-26 2017-08-17 Suntory Holdings Limited Method for producing hop extract
JP7011585B2 (ja) * 2015-11-03 2022-01-26 ピューラック バイオケム ベスローテン フェンノートシャップ キサントフモールを含む抗微生物剤および食品における使用
CN105687319A (zh) * 2016-02-23 2016-06-22 中国人民解放军第二军医大学 一种抗骨质疏松啤酒花提取物及其制备方法和应用
CN106198785B (zh) * 2016-06-27 2019-12-03 广州纤维产品检测研究院 一种超临界co2快速分离纺织品中阻燃剂的方法
CN106290667A (zh) * 2016-08-24 2017-01-04 广州纤维产品检测研究院 采用超临界co2流体快速分离纺织品中有机氯农药的方法
US20180028436A1 (en) * 2016-10-15 2018-02-01 Fatemeh Abdi Topical formulation of hops extract
CN108017604B (zh) * 2016-11-02 2021-05-07 沈阳药科大学 α-酸衍生物及其制备方法和用途
JP2018161108A (ja) * 2017-03-27 2018-10-18 キリン株式会社 脂肪酸エステル含有ホップ組成物の製造方法
PL242648B1 (pl) * 2017-06-30 2023-04-03 Bioactive Tech Spolka Z Ograniczona Odpowiedzialnoscia Sposób otrzymywania ksantohumolu
IT201800005721A1 (it) * 2018-05-25 2019-11-25 Nuove composizioni per il trattamento di leiomiomi uterini
JPWO2020031952A1 (ja) * 2018-08-10 2021-08-26 サントリーホールディングス株式会社 血糖値上昇抑制用組成物及び血糖値上昇抑制方法
CN109329595A (zh) * 2018-10-29 2019-02-15 齐鲁工业大学 一种酒花制品替代抗生素在饲料添加剂上的应用
CN109796319B (zh) * 2019-01-31 2021-11-23 广西医科大学附属肿瘤医院 异戊烯基查尔酮衍生物的合成方法及其在制药中的应用
JPWO2021131570A1 (fr) * 2019-12-25 2021-07-01
CN115650942B (zh) * 2022-10-25 2024-06-04 长沙市惠瑞生物科技有限公司 从啤酒花中提取分离纯化8-异戊烯基柚皮素的方法
CN116036153B (zh) * 2023-02-22 2023-09-26 深圳市儿童医院 啤酒花提取物在制备用于治疗或者预防抑郁症、抑郁症并发症、焦虑症的产品中的应用

Family Cites Families (7)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
DE2244065A1 (de) * 1972-09-08 1974-03-14 Haarmann & Reimer Gmbh Verfahren zur extraktion von hopfen
CH649778A5 (de) * 1978-06-20 1985-06-14 Mueller Adam Verfahren zum isomerisieren von alpha-saeuren in einem hopfenextrakt.
DE3103617A1 (de) * 1981-02-03 1982-08-05 Horst & Klotz Hopfenveredelung GmbH, 8302 Mainburg Verfahren zur extraktion von bitterstoffen und gerbstoffen aus hopfen
US4507329A (en) * 1982-02-03 1985-03-26 S. S. Steiner, Inc. Mixed solvent extraction of hops
DE3632401A1 (de) * 1986-09-24 1988-04-07 Barth Raiser Hopfenextraktion Verfahren zur extraktion unpolarer inhaltsstoffe aus hopfen
US5972411A (en) * 1997-04-03 1999-10-26 Miller Brewing Company Methods of making and using purified kettle hop flavorants
DE19939350B4 (de) * 1999-08-19 2005-04-07 Plantextrakt Gmbh & Co. Kg Hopfenextrakt und Verfahren zu dessen Herstellung

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ATE297460T1 (de) 2005-06-15
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CN1269942C (zh) 2006-08-16
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DE50203359D1 (de) 2005-07-14
DE10139479A1 (de) 2003-02-27
MXPA04001226A (es) 2005-06-20
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US20050042318A1 (en) 2005-02-24
ES2240787T3 (es) 2005-10-16

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